DNA construct and in vitro test for detecting tumor promoters by means of said DNA construct

ABSTRACT

The invention relates to a DNA construct and to a method of detecting tumor promoters by employing this DNA construct. 
     It is an object of the invention to make available an easily implemented quantitative detection method for tumor promoters. 
     This is accomplished by a DNA construct including at least one reporter gene and a control region for the activation of a particularly strongly induced gene of the Epstein-Barr virus (EBV), and by the action of the substance to be examined on EBV-containing cells that contain the DNA construct, whereupon the resulting quantity of reporter gene product is quantitatively determined.

The invention relates to a DNA construct and to a method of detecting tumor promoters with the use of this DNA construct.

Tumor promoters are substances which, in combination with doses of solitary carcinogens that are in themselves not tumorigenic, induce the development of tumors or carcinomas. Solitary carcinogens are substances which produce tumors when applied individually (in animal tests) in dependence on the dosage. However, the sole influence of tumor promoters does not produce cancer in the sense of their definition, but it enhances to a considerable degree the development of tumors in organs which have been initiated by the exogenous influence of a solitary carcinogen. Simple and quickly performed tests systems are already available to identify solitary carcinogens. However, in the past, the substance class of tumor promoters could be identified only by way of animal experiments, for example, on the skin of a mouse or the intestines of a rat.

A starting procedure for a test to search out tumor promoters described by H. zur-Hausen, G. W. Bornkamm, R. Schmidt, and E. Hecker, entitled "Tumor Initiators and Promoters in the Induction of Epstein-Barr Virus", in Proceedings of the National Academic Science [sic] (U.S.A.), Volume 76, pages 782-785 (1979), provides for a determination of the property of substances that induces the Epstein-Barr virus (EBV) by means of an immunofluorescence display of EBV-coded antigens (so-called early antigens). However, this technique requires personnel that are trained in virology and serology procedures. Moreover, this process is time consuming and, due to the subjective evaluation of immunofluorescence stained cells, not accurately standardizable.

The final proof of the tumor promoting property of a substance can be brought only by the initiation of tumors in an in-vivo animal experiment, for example, on the skin of a mouse. However, such in-vivo experiments are not suitable as search tests for testing a plurality of substances.

It is now an object of the invention to make available a DNA construct and an easily performed quantitative method of the above-mentioned type.

This is accomplished by claims 1 and 8. The remaining claims define advantageous features of the invention.

The invention will now be described in greater detail with reference to two embodiments thereof and FIGS. 1 to 5.

FIG. 1 is a schematic representation of the configuration of the DNA construct,

while FIGS. 2 and 3 are schematic representations of two DNA constructs with points of intersection and their precise positions.

FIG. 4 shows a Southern-blot analysis of various pHEBO-DR-CAT transfectants (A4, D2, D4), and

FIG. 5 depicts a CAT assay of the pHEBO-DR-CAT transfected Raji sublines D4, A4 and D2.

The base sequence (SEQ ID NOS: 1 and 2) and mapping of these two DNA constructs are given in Tables 1 and 2.

The invention is based on the observation that different classes of tumor promoters, such as diterpene esters, indole alkaloids or polyacetates, Teleocidin, transforming growth factor β (TGFβ), induce a lytic or abortive virus cycle in lymphatic cells containing the Epstein-Barr virus (EBV). The EBV genome contains a large number of genes that are not expressed in the latent state and are induced only upon the induction of a lytic or abortive cycle. The control region of the particularly strongly induced DR gene responsible for this activation (DL or others are possible) was cloned by molecular biology methods ahead of the coding region of a so-called reporter gene. The vector (pHEBO) employed for this purpose carries the episomal replication origin of EBV and two resistance genes one of which permits the selection of transfected eukaryotic cells with hygromycin and the other the selection in procaryotes.. This vector is described by B. Sugden, K. Marsh and J. A. Yates as "[A] vector that replicates as a plasmid and can be efficiently selected in B-lymphoblasts transformed by Epstein virus" in Molecular and Cellular Biology, Volume 5, pages 410-413 (1985). The construct obtained in this way (see also FIG. 1) was introduced in stable form into a cell line (Raji) containing EBV.

Raji cells were selected since, due to deletions, no infectious virus can be formed in the Raji-EBV genome, and because the so-called spontaneous induction rate in these cells is very low. Target cells for the stable transfection with the above-described plasmid can be all EBV positive cell lines which can be induced by chemical inductors (e.g., the cell line P3HR1 or Akata). The translation product of the reporter gene can be detected quantitatively in a relatively simple manner by way of an enzyme reaction in extracts of these cells. The quantity of detectable reporter gene product correlates quantitatively with the activation state of the EBV genome.

Due to the good correlation between tumor promoting and EBV inducing activity, this system can be employed as an easily and quickly performed search test for the identification of tumor promoters.

Tested were the reporter genes (REP) chloramphenicol acetyltransferase (CAT) and firefly luciferase (LUC).

Other usable reporter genes are: β-galactosidase, the β-globin gene, hepatitis B coded antigens, or the like.

The test system described here differs from the prior art systems primarily by its simplified implementation and a decisive improvement in the quantitative evaluation of the results. In the past, the subjective evaluation of immunofluorescence stained cells was included in each measuring result. In particular, the statement that a certain percentage of cells exhibits positive immunofluorescence for early EBV antigens includes a subjective determination of a threshold value. Moreover, the absolute protein quantity of early antigens developed in the individual cell cannot be determined in this way. In the past this made standardization considerably more difficult.

The quantity of resulting reporter gene product, however, can be determined accurately and reproducibly in a given number of cells. Another advantage of this system is the clonal origin of the stably transfected Raji-DR-REP line. It is known that during the long in-vitro cultivation of the Raji cell line, subpopulations are created which exhibit a different biological behavior. In addition, this test is distinguished by greater sensitivity and an improved dosage to effect ratio.

FIG. 1 is a diagram of the construction of the DNA clones (DNA constructs) DR-REP and pHEBO-DR-REP.

REP stands for reporter gene,

The top line in the drawing represents the genome of the EBV in linear form (terminal repetitions, TR; internal repetitions, IR; duplicated region left and right, DL and DR; long and short uniform region, U_(L) and U_(S)). The next line is an enlarged representation of the BglII restriction fragment (BglII-K) (Polack et al, "A complete Set of Overlapping Cosmid Clones of M-ABA Virus Derived From Nasopharyngeal Carcinoma and Its Similarity to Other Epstein-Barr Virus Isolates" in Genes, Volume 27, pages 279-288, 1984), which carries the DR gene. The arrow in the next line corresponds to the messenger RNS of a size of 2.8 kb which was coded by this region.

The construction of pHEBO-DR-CAT with chloramphenicol acetyltransferase (CAT) as the reporter gene will now be described. The promoter region of the DR gene was cloned as a HindIII-NdeI fragment (after the conversion of the BglI point of intersection, which was disposed 3' away from the DR promoter, into a HindIII point of intersection) from pMBgllII-K ahead of the CAT reporter gene at the points of intersection HindIII/Ndel of the pSV2CAT vector (the SV40 promoter in the pSV2CAT was thus deleted).

The DR-promoter-CAT component of the DR-CAT clone was inserted as an XbaI-BamHI fragment at the points of intersection BamH I and XbaI of the puc18(R/X) vector (laboratory identification: G3-28). The puc18(R/X) vector developed from the conversion of the EcoR I point of intersection into an XhoI point of intersection in vector puc18. From Ge-28, DR-CAT was mobilized as an SphI-XhoI fragment and cloned into the SphI/SalI point of intersection of vector pHEBO (pHEBO-DR-CAT).

The promoter of the DR gene is identified as a black box. The point of initiation of polymerase II is represented by a short arrow oriented toward the left. The SV40 component, 3' removed from the open reading frame of the CAT gene, is shown in hatching. The term ori-P represents the "origin of plasmid replication" of EBV, HYG stands for the hygromycin resistance gene and AMP for the resistance gene against ampicillin.

FIG. 4 shows a Southern-blot analysis of various pHEBO-DR-CAT transfectants (A4, D2, D4).

In each case, approximately 10 μg of cellular DNA of sublines A4, D2 and D4 were applied either unsplit (U) or split with BglII (B) or HindIII (H), onto an 0.6% agarose gel. The unsplit (U) and the HindIII (H) split plasmid pHEBO-CR-CAT, respectively, served as a control, of which a one-fold (1×), five-fold (5×) and ten-fold (10×) amount of a genome equivalent was applied. After the gel run, the DNA, which was separated according to size, was transferred to nitrocellulose filters and hybridized with ³² P-marked DNA of pHEBO-DR-CAT. The cloned pHEBO-DR-CAT contains sequences which hybridize with the oriP as well as with EBV regions DL and DR.

In the cellular DNAs (A4, D2, D4) cut with HindIII, there appear five bands in the autoradiogram: the HindIII fragments A, B and D2 of the endogenous Raji-EBV genome, and two fragments of a size of 8 and 2.5 kb corresponding to the transfected DNA (marked x; see right margin). The BglII fragments A, C and K of the endogenous Raji EBV genome are represented through Bgl II digestion. Since Bgl II does not intersect in the cloned pHEBO-DR-CAT, the identical band pattern as in the unsplit applied plasmid (pHEBO-DR-CAT: U) results for the transfected DNA (marked with an x at the left margin of the autoradiogram).

The result of this Southern-blot analysis leads to the conclusion that the transfected plasmid is contained in the episomal state and in about 50 to 100 copies in all three sublines.

A simplified restriction enzyme map of the EBV genome is represented below the autoradiogram.

FIG. 5 depicts a CAT assay of the pHEBO-DR-CAT transfected Raji sublines D3, A4 and D2.

Approximately 20 to 30×10⁶ cells were treated for 48 hours with the tumor promoter TPA (20 ng/ml sigma) and 5-iodo-2'desoxyuridin (50 μg/ml sigma) as intensifier. The CAT enzyme developed only in the induced cells (T+I) as indicated by the occurrence of acetylized forms (marked with 2, 3, 4 in the right margin) of the ¹⁴ C-marked chloramphenicol in the enzyme assay. No CAT activity can be detected in untreated cells (U).

The detection of tumor promoting properties of compounds (detection of tumor promoters) for both variations is effected according to the following schemes.

(a) Raji Cells Transfected with pHEBO-DR-CAT

(1) growing at least 1 to 5×10⁶ transfected cells per experiment;

cell culture conditions:

cell culture medium: RPMI 1640 (Gibco) with: approximately 10% fetal calf serum, 300 μg/ml hygromycin (Calbiochem), 100 u/ml penicillin, 280 μg/ml L-glutamine, approximately 37° C., 5 to 6% CO₂.

(2) incubation of these cells with a corresponding quantity of the substance to be tested for 48 hours (=standard starting mixture; <20 hours up to several days are also possible) at 37° C. and in 6% CO₂ ;

(3) recovery of the cells by centrifuging;

(4) resuspension of the cells in 100 μl 0.25M tris-HCl (pH 7.8);

(5) break-up of the cells to obtain the developed enzyme by means of ultrasound treatment or three cycles of freezing/thawing (3 minutes in a CO₂ /ethanol bath/3 minutes thawing at 30° C.);

(6) clarification the cell lysate by centrifuging;

(7) determination of the protein concentration;

(8) incubation of, for example,

20 μl lysate (quantity depends on the protein quantity);

100 μl 0.25M tris-HCl (pH 7.8);

1 μCi ¹⁴ C-chloramphenicol;

20 μl 4 mM acetyl coenzyme A;

(9) incubation at 37° C. for one hour (longer incubation may increase sensitivity somewhat);

(10) interruption of the enzyme reaction by the addition of 2 ml cold ethyl acetate;

(11) intensive mixing of the reaction mixture for extraction of the chloramphenicol and the developed derivatives;

(12) centrifugation in order to separate the phases;

(13) removal of the organic phase;

(14) inspissation of the organic phase from (13) by applying a vacuum with simultaneous rotation;

(15) resuspension of the remaining solid phases from (14) in 30 μl ethyl acetate;

(16) thin film chromatography with chloroform ethanol (95:5) as the running agent for the separation of the different acetylized forms of chloramphenicol on silica gel plates (Polygram Sil G, Macherey-Nagel);

(17) autoradiography of the thin-film chromatography plates or evaluation with the aid of a radioactivity scanner.

(b) Raji Cells Transfected with pHEBO-DR-LUC

(1) growing at least 0.1 to 1.0×10⁴ transfected cells per experiment;

cell culture conditions:

cell culture medium: RPMI 1640 (Gibco) with: 10% fetal calf serum, 300 μg/ml hygromycin (Calbiochem), 100 u/ml penicillin, 280 μg/ml L-glutamine, 37° C., 5 to 6% CO₂ ;

(2) incubation of these cells with a corresponding quantity of substance to be tested for 48 hours (=standard starting mixture; <20 hours up to several days is also possible) at 37° C. and in 6% CO₂ ;

(3) recovery of the cells by centrifuging;

(4) resuspension of the cells in lyse buffer (100 mM potassium phosphate, ph 7.8, 1 mM DTT);

(5) break-up of the cells to obtain the developed enzyme by means of ultrasound treatment of three cycles of freezing/thawing (3 minutes in a CO₂ /ethanol bath/3 minutes thawing at 30° C.);

(6) clarification of the cell lysate by centrifuging;

(7) determination of the protein concentration;

(8) incubation of, for example,

20 μl lysate (quantity depends on the protein quantity);

100 μl test buffer (25 mM glycyl glycine, pH 7.8; 5 mMATP, 15 mMMgSo₄);

injection of

100 μl luciferin solution (1 mM luciferin in 0.5M tris-Hcl (pH 7.8);

(9) immediate measurement of the emitted light in a luminometer (e.g., Lumat by Berthold).

The quantity of the detected enzymes is a measure for the EBV-inducing property of the examined substances and thus permits a conclusion as to the possible tumor promoting property of this substance.

The particular advantages of method (b) over method (a) are the significantly lower quantity of cells, a sensitivity that is higher by a multiple, a significantly simplified quantifiability (the quantity of resulting enzyme can be quantitatively determined in a luciferase test through several logging stages without additional dilution series), a noticeably lower amount of work involved and the avoidance of radioactive waste.

                                      TABLE 1                                      __________________________________________________________________________     pHEBO-DR-CAT                                                                   __________________________________________________________________________     Features:                                                                      Identification in record book:                                                                 DRCATPHEBO                                                     Cloning:        DR-CAT as XhoI-SpHI 3.6 kb fragment cloned into SalI-SpHI                      of pHEBO                                                       Total bases:    10,580                                                         Restriction Enzyme Intersections:                                                              Position:                                                      __________________________________________________________________________     HindIII         29                                                             BamHI           375                                                            SphI            562                                                            SalI            574                                                            HindIII         2,539                                                          EcoRI           2,789                                                          BamHI           4,171                                                          EcoRI           7,895                                                          EcoRI           8,292                                                          EcoRI           10,578                                                         __________________________________________________________________________     Regions:        Position:                                                      __________________________________________________________________________     DR              575-2,539                                                      CAT             2,539-3,325                                                    SV40            3,325-4,171                                                    EBV-oriP        4,603-6,783                                                    hygromycin resistance gene                                                                     8,383-6,783                                                    ampicillin resistance gene                                                                     10,333-9,483                                                   __________________________________________________________________________     Sequence (SEQ ID NO: 1):                                                       TTCTCATGTTTGACAGCTTATCATCGATAAGCTTTAATGCGGTAGTTTATCACAGTTAAATT                 GCTAACGCAGTCAGGCACCGTGTATGAAATCTAACAATGCGCTCATCGTCATCCTCGGCACC                 GTCACCCTGGATGCTGTAGGCATAGGCTTGGTTATGCCGGTACTGCCGGGCCTCTTGCGGGA                 TATCGTCCATTCCGACAGCATCGCCAGTCACTATGGCGTGCTGCTAGCGCTATATGCGTTGA                 TGCAATTTCTATGCGCACCCGTTCTCGGAGCACTGTCCGACCGCTTTGGCCGCCGCCCAGTC                 CTGCTCGCTTCGCTACTTGGAGCCACTATCGACTACGCGATCATGGCGACCACACCCGTCCT                 GTGGATCCTCTACGCCGGACGCATCGTGGCCGGCATCACCGGCGCCACAGGTGCGGTTGCTG                 GCGCCTATATCGCCGACATCACCGATGGGGAAGATCGGGCTCGCCACTTCGGGCTCATGAGC                 GCTTGTTTCGGCGTGGGTATGGTGGCAGGCCCCGTGGCCGGGGGACTGTTGGGCGCCATCTC                 CTTGCATGCCTGCAGGTCGACTCTAGAACTGGATACTCGTGGGGGGTAAAGAAGGTGAATAA                 AAATTACAAACATTCTCTGCCCAGCCTTCAGACTATCAAACCTAGATAATCATTCTATAAAT                 AACCCAGGGCTTGCATCACAATTTAAAGGGAAGTTAAGATGGGGGATTACCTTCTAAGAGGA                 AATGACAAGCCACAGACATCCCCACATATAGCCGGGTGTCCACTGTCTAATGTCAGTAATTT                 AATTCCATAGTGAAAATAGCACCCCCAACTCAATTTGGAATCCAGAAACTATATTGCACACC                 AACACCCCCTCCTCTGCACATGAGCAAGACAAGACATCTATGTTTATCTCTAAATGTGCCAT                 GGAACCCGGTTGCCCATGCAGTGGTGTCAGACAGGAAAATGGTTAATTAACCACATCTTAAA                 TTGCCACCTTAGGCAAATTGAAAATAGGCAGGCTGAAGAATTGCACAATACCCTAAACAGGT                 AAGAGGAATTAGTGACATTTTATGAATTTTTTTACAAACTTTCACACTCAAGAATAGAAACC                 CAATACCAACAGGTGTGCAGGTGTGCATGACAAATCTTGGGGGTCTCAGAACCCAGGACCAG                 ACTTTGAAGTCTCAGGTATAGGTCCTGGCTGAGATTCTATTAATAAAACAAGAGAGAAAGAA                 GGCGGGCGCCCATTAGAATCTGCTCGGCTGCCAGTAAGTTGCCAGCAAACAGGAACACAAAC                 AAACCAAGGGTGTTGGCCCCTACAGGCTCCCAAGGCGGGGGTTGGGCACAGGGCCAAGCTCT                 GCCACCACAGGAGGCAAGTAGACATGCAGGAACACATGGCCCTGGCTAGGAAAGGGAGGAAA                 TAGAGGCCACAGCCAAAGTTAGGCTGCCGCCCCACCTGTGTACCCAGGGTGAGAGACCTTGG                 GAGGTCGTCAGCTTAACCAGCGCCGCCCTCACCCCATTGCCAACTTCCGGCTCACACAAAAC                 CACTCCCAAAAATTGAAGACTGGCCAAAATCCAGCTTCCGTCCCCGGGACGTGGTGCTTCCT                 AAAGGCGGGGCTCATGGATTAGCAGGGGCTTAGTGTGTCATGGTGAGGCAGGCAAGGCGAGC                 AACGGGGGCTTAGTGGCTCAAAGTGATGCATCCCAAAGGCAGCCACCACGCTGGAGGGACAT                 TGTCCACGGGACAAGGCACAGGCCAGGTCATGACCCAGGAAGTGGCGAGCATCGGTCAGCTG                 ACCAAATGTGCAAAGGTGACAAGTCAGTAAGGCACGCGGGGGGCCACGTCACCCCGGGGTGC                 TGGGGTGGGGGATGGGCTCAGGCAACCGTAAGGGAGGGGGGGGTAGGGGGGGGAGGGATTAC                 ACTATAGGGTTCCCTTCCTCTAGGTTCTATATACCTATAGGTATATACCCAGCTGCAATACC                 CTATTCCACCACTAGGTTAATAACCTATAGGTTATTCTACCATTAAAACGGAAGGAGGAAGG                 GTGGCGCACCTTAAGGTAGGGTAGGGGGGTACCCCAGTAGGAACCTAGCTGAATCCTACCTA                 GCTCCACCCACCTGGTATATAGGGGCGGAGCTTAGGATACCTCCAGGATAATGGAACCCTAT                 GGAGACCTACCTCTAGGCTCCACCCACTAGGTATATCGGGGCGGAGCCCACTCCTCCCCCTC                 CTGGTTCAACCCTATGGAGGGGACCCTCCTGAGGCTCCGCCTACCCCAAATCTCGCGGGCCT                 CTAGCCCCTCCTCCTCTCGTTATCCCAATAGAATGACCTCCAGGTACCACCCACCTGGTTAC                 ACACCTTAATGTAACCCAACGGGCTAAAATCACACACCTGAATTAACCAATGAGAAGCCCCC                 CACACCTGAGCAAACCTTAAGGTATTGCACAGAAACCCCAAAAAGAGGATAAAAGAAGGCGA                 GCTGGCCCGGCTCGCCAGCGTCGTCCAGACGCTCGGGGGGTGCACACCTCCCAGCCGCAAGC                 TTGGCGAGATTTTCAGGAGCTAAGGAAGCTAAAATGGAGAAAAAAATCACTGGATATACCAC                 CGTTGATATATCCCAATGGCATCGTAAAGAACATTTTGAGGCATTTCAGTCAGTTGCTCAAT                 GTACCTATAACCAGACCGTTCAGCTGGATATTACGGCCTTTTTAAAGACCGTAAAGAAAAAT                 AAGCACAAGTTTTATCCGGCCTTTATTCACATTCTTGCCCGCCTGATGAATGCTCATCCGGA                 ATTCCGTATGGCAATGAAAGACGGTGAGCTGGTGATATGGGATAGTGTTCACCCTTGTTACA                 CCGTTTTCCATGAGCAAACTGAAACGTTTTCATCGCTCTGGAGTGAATACCACGACGATTTC                 CGGCAGTTTCTACACATATATTCGCAAGATGTGGCGTGTTACGGTGAAAACCTGGCCTATTT                 CCCTAAAGGGTTTATTGAGAATATGTTTTTCGTCTCAGCCAATCCCTGGGTGAGTTTCACCA                 GTTTTGATTTAAACGTGGCCAATATGGACAACTTCTTCGCCCCCGTTTTCACCATGGGCAAA                 TATTATACGCAAGGCGACAAGGTGCTGATGCCGCTGGCGATTCAGGTTCATCATGCCGTCTG                 TGATGGCTTCCATGTCGGCAGAATGCTTAATGAATTACAACAGTACTGCGATGAGTGGCAGG                 GCGGGGCGTAATTTTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAA                 TAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTT                 CTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAA                 AATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCA                 ACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTG                 CTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTC                 CAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTG                 AGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAA                 AGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATA                 ACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATT                 AATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGA                 ATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTT                 TTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAAT                 TGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAA                 ATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAAT                 GTATCTTATCATGTCTGGATCCCCGGGTACCGAGCTCGCCTCGACCGATGCCCTTGAGAGCC                 TTCAACCCAGTCAGCTCCTTCCGGTGGGCGCGGGGCATGACTATCGTCGCCGCACTTATGAC                 TGTCTTCTTTATCATGCAACTCGTAGGACAGGTGCCCTGGCCGGGGTCCCGCGGAAACTCGG                 CCGTGGTGACAGGAAAAGGACAAGCAGCGAAAATTCACGCCCCCTTGGGAGGTGGCGGCATA                 TGCAAAGGATAGCACTCCCACTCTACTACTGGGTATCATATGCTGACTGTATATGCATGAGG                 ATAGCATATGCTACCCGGATACAGATTAGGATAGCATATACTACCCAGATATAGATTAGGAT                 AGCATATGCTACCCAGATATAGATTAGGATAGCCTATGCTACCCAGATATAAATTAGGATAG                 CATATACTACCCAGATATAGATTAGGATAGCATATGCTACCCAGATATAGATTAGGATAGCC                 TATGCTACCCAGATATAGATTAGGATAGCATATGCTACCCAGATATAGATTAGGATAGCATA                 TGCTATCCAGATATTTGGGTAGTATATGCTACCCAGATATAAATTAGGATAGCATATACTAC                 CCTAATCTCTATTAGGATAGCATATGCTACCCGGATACAGATTAGGATAGCATATACTACCC                 AGATATAGATTAGGATAGCATATGCTACCCAGATATAGATTAGGATAGCCTATGCTACCCAG                 ATATAAATTAGGATAGCATATACTACCCAGATATAGATTAGGATAGCATATGCTACCCAGAT                 ATAGATTAGGATAGCCTATGCTACCCAGATATAGATTAGGATAGCATATGCTATCCAGATAT                 TTGGGTAGTATATGCTACCCATGGCAACATTAGCCCACCGTGCTCTCAGCGACCTCGTGAAT                 ATGAGGACCAACAACCCTGTGCTTGGCGCTCAGGCGCAAGTGTGTGTAATTTGTCCTCCAGA                 TCGCAGCAATCGCGCCCCTATCTTGGCCCGCCCACCTACTTATGCAGGTATTCCCCGGGGTG                 CCATTAGTGGTTTTGTGGGCAAGTGGTTTGACCGCAGTGGTTAGCGGGGTTACAATCAGCCA                 AGTTATTACACCCTTATTTTACAGTCCAAAACCGCAGGGCGGCGTGTGGGGGCTGACGCGTG                 CCCCCACTCCACAATTTCAAAAAAAAGAGTGGCCACTTGTCTTTGTTTATGGGCCCCATTGG                 CGTGGAGCCCCGTTTAATTTTCGGGGGTGTTAGAGACAACCAGTGGAGTCCGCTGCTGTCGG                 CGTCCACTCTCTTTCCCCTTGTTACAAATAGAGTGTAACAACATGGTTCACCTGTCTTGGTC                 CCTGCCTGGGACACATCTTAATAACCCCAGTATCATATTGCACTAGGATTATGTGTTGCCCA                 TAGCCATAAATTCGTGTGAGATGGACATCCAGTCTTTACGGCTTGTCCCCACCCCATGGATT                 TCTATTGTTAAAGATATTCAGAATGTTTCATTCCTACACTAGTATTTATTGCCCAAGGGGTT                 TGTGAGGGTTATATTGGTGTCATAGCACAATGCCACCACTGAACCCCCCGTCCAAATTTTAT                 TCTGGGGGCGTCACCTGAAACCTTGTTTTCGAGCACCTCACATACACCTTACTGTTCACAAC                 TCAGCAGTTATTCTATTAGCTAAACGAAGGAGAATGAAGAAGCAGGCGAAGATTCAGGAGAG                 TTCACTGCCCGCTCCTTGATCTTCAGCCACTGCCCTTGTGACTAAAATGGTTCACTACCCTC                 GTGGAATCCTGACCCCATGTAAATAAAACCGTGACAGCTCATGGGGTGGGAGATATCGCTGT                 TCCTTAGGACCCTTTTACTAACCCTAATTCGATAGCATATGCTTCCCGTTGGGTAACATATG                 CTATTGAATTAGGGTTAGTCTGGATAGTATATACTACTACCCGGGAAGCATATGCTACCCGT                 TTAGGGTTAACAAGGGGGCCTTATAAACACTATTGCTAATGCCCTCTTGAGGGTCCGCTTAT                 CGGTAGCTACACAGGCCCCTCTGATTGACGTTGGTGTAGCCTCCCGTAGTCTTCCTGGGCCC                 CTGGGAGGTACATGTCCCCCAGCATTGGTGTAAGAGCTTCAGCCAAGAGTTACACATAAAGG                 CAATGTTGTGTTGCAGTCCACAGACTGCAAAGTCTGCTCCAGGATGAAAGCCACTCAGTGTT                 GGCAAATGTGCACATCCATTTATAAGGATGTCAACTACAGTCAGAGAACCCCTTTGTGTTTG                 GTCCCCCCCCGTGTCACATGTGGAACAGGGCCCAGTTGGCAAGTTGTACCAACCAACTGAAG                 GGATTACATGCACTGCCCCGTGACCAATACAAAACAAAAGCGCTCCTCGTACCAGCGAAGAA                 GGGGCAGAGATGCCGTAGTCAGGTTTAGTTCGTCCGGCGGCGCCAGAAATCCGCGCGGTGGT                 TTTTGGGGGTCGGGGGTGTTTGGCAGCCACAGACGCCCGGTGTTCGTGTCGCGCCAGTACAT                 GCGGTCCATGCCCAGGCCATCCAAAAACCATGGGTCTGTCTGCTCAGTCCAGTCGTGGACCT                 GACCCCACGCAACGCCCAAAAGAATAACCCCCACGAACCATAAACCATTCCCCATGGGGGAC                 CCCGTCCCTAACCCACGGGGCCCGTGGCTATGGCGGGCTTGCCGCCCCGACGTTGGCTGCGA                 GCCCTGGGCCTTCACCCGAACTTGGGGGTTGGGGTGGGGAAAAGGAAGAAACGCGGGCGTAT                 TGGCCCCAATGGGGTCTCGGTGGGGTATCGACAGAGTGCCAGCCCTGGGACCGAACCCCGCG                 TTTATGAACAAACGACCCAACACCCGTGCGTTTTATTCTGTCTTTTTATTGCCGTCATAGCG                 CGGGTTCCTTCCGGTATTGTCTCCTTCCGTGTTTCAGTTAGCCTCCCCCATCTCCCGATCCC                 CTCGGACGAGTGCTGGGGCGTCGGTTTCCACTATCGGCGAGTACTTCTACACAGCCATCGGT                 CCAGACGGCCGCGCTTCTGCGGGCGATTTGTGTACGCCCGACAGTCCCGGCTCCGGATCGGA                 CGATTGCGTCGCATCGACCCTGCGCCCAAGCTGCATCATCGAAATTGCCGTCAACCAAGCTC                 TGATAGAGTTGGTCAAGACCAATGCGGAGCATATACGCCCGGAGCCGCGGCGATCCTGCAAG                 CTCCGGATGCCTCCGCTCGAAGTAGCGCGTCTGCTGCTCCATACAAGCCAACCACGGCCTCC                 AGAAGAAGATGTTGGCGACCTCGTATTGGGAATCCCCGAACATCGCCTCGCTCCAGTCAATG                 ACCGCTGTTATGCGGCCATTGTCCGTCAGGACATTGTTGGAGCCGAAATCCGCGTGCACGAG                 GTGCCGGACTTCGGGGCAGTCCTCGGCCCAAAGCATCAGCTCATCGAGAGCCTGCGCGACGG                 ACGCACTGACGGTGTCGTCCATCACAGTTTGCCAGTGATACACATGGGGATCAGCAATCGCG                 CATATGAAATCACGCCATGTAGTGTATTGACCGATTCCTTGCGGTCCGAATGGGCCGAACCC                 GCTCGTCTGGCTAAGATCGGCCGCAGCGATCGCATCCATGGCCTCCGCGACCGGCTGCAGAA                 CAGCGGGCAGTTCGGTTTCAGGCAGGTCTTGCAACGTGACACCCTGTGCACGGCGGGAGATG                 CAATAGGTCAGGCTCTCGCTGAATTCCCCAATGTCAAGCACTTCCGGAATCGGGAGCGCGGC                 CGATGCAAAGTGCCGATAAACATAACGATCTTTGTAGAAACCATCGGCGCAGCTATTTACCC                 GCAGGACATATCCACGCCCTCCTACATCGAAGCTGAAAGCACGAGATTCTTCGCCCTCCGAG                 AGCTGCATCAGGTCGGAGACGCTGTCGAACTTTTCGATCAGAAACTTCTCGACAGACGTCGC                 GGTGAGTTCAGGCTTTTTCATATCTCATTGCCCCCGGGGGATCTGCGGCACGCTGTTGACGC                 TGTTAAGCGGGTCGCTGCAGGGTCGCTCGGTGTTCGAGGCCACACGCGTCACCTTAATATGC                 GAAGTGGACCTGGGACCGCGCCGCCCCGACTGCATCTGCGTGTTCGAATTCGCCAATGACAA                 GACGCTGGGCGGGGTTTGTGTCATCATAGAACTAAAGACATGCAAATATATTTCTTCCGGGG                 ACACCGCCAGCAAACGCGAGCAACGGGCCACGGGGATGAAGCAGGGCGGCACCTCGCTAACG                 GATTCACCACTCCAAGAATTGGAGCCAATCAATTCTTGCGGAGAACTGTGAATGCGCAAACC                 AACCCTTGGCAGAACATATCCATCGCGTCCGCCATCTCCAGCAGCCGCACGCGGCGCATCTC                 GGGCAGGCCGACGCGCTGGGCTACGTCTTGCTGGCGTTCGCGACGCGAGGCTGGATGGCCTT                 CCCCATTATGATTCTTCTCGCTTCCGGCGGCATCGGGATGCCCGCGTTGCAGGCCATGCTGT                 CCAGGCAGGTAGATGACGACCATCAGGGCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGT                 TGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGT                 CAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCT                 CGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGG                 GAAGCGTGGCGCTTTCTCAATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGC                 TCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAA                 CTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTA                 ACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAAC                 TACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGG                 AAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTG                 TTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCT                 ACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATC                 AAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTA                 TATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCG                 ATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACG                 GGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTC                 CAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACT                 TTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGT                 TAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTG                 GTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTG                 TGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGT                 GTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGAT                 GCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCG                 AGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGT                 GCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGAT                 CCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGC                 GTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACG                 GAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATT                 GTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGC                 ACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTA                 TAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCAAGAA                                       __________________________________________________________________________

                                      TABLE 2                                      __________________________________________________________________________     pHEBO-DR-LUC                                                                   __________________________________________________________________________     Features:                                                                      Identification in record book:                                                                    RF202-15                                                    Cloning: DR-LUC cloned as BamHI+BglII 4.54 kb fragment into                    BAMHI of pHEBO.                                                                Total bases:       11,616                                                      Restriction Enzyme Intersections:                                                                 Position:                                                   __________________________________________________________________________     BamHI/BglII (Sau3A)                                                                               1                                                           HindIII            1,964                                                       EcoRI              2,580                                                       BamHI              4,547                                                       EcoRI              8,556                                                       EcoRI              8,953                                                       EcoRI              11,239                                                      HindIII            11,270                                                      __________________________________________________________________________     Regions:           Positions:                                                  __________________________________________________________________________     DR (of EBV)        1-1,963                                                     Luciferase (LUC)   1,972-3,689                                                 part of SV40       3,701-4,546                                                 EBV-oriP (of EBV)  4,979-7,159                                                 hygromycin resistance gene                                                                        9,044-7,159                                                 ampicillin res. gene (AMP) (of pBR322)                                                            10,994-10,144                                               __________________________________________________________________________     Sequence (SEQ ID NO: 2):                                                       GATCTCTAGAACTGGATACTCGTGGGGGGTAAAGAAGGTGAATAAAAATTACAAACATTCTC                 TGCCCAGCCTTCAGACTATCAAACCTAGATAATCATTCTATAAATAACCCAGGGCTTGCATC                 ACAATTTAAAGGGAAGTTAAGATGGGGGATTACCTTCTAAGAGGAAATGACAAGCCACAGAC                 ATCCCCACATATAGCCGGGTGTCCACTGTCTAATGTCAGTAATTTAATTCCATAGTGAAAAT                 AGCACCCCCAACTCAATTTGGAATCCAGAAACTATATTGCACACCAACACCCCCTCCTCTGC                 ACATGAGCAAGACAAGACATCTATGTTTATCTCTAAATGTGCCATGGAACCCGGTTGCCCAT                 GCAGTGGTGTCAGACAGGAAAATGGTTAATTAACCACATCTTAAATTGCCACCTTAGGCAAA                 TTGAAAATAGGCAGGCTGAAGAATTGCACAATACCCTAAACAGGTAAGAGGAATTAGTGACA                 TTTTATGAATTTTTTTACAAACTTTCACACTCAAGAATAGAAACCCAATACCAACAGGTGTG                 CAGGTGTGCATGACAAATCTTGGGGGTCTCAGAACCCAGGACCAGACTTTGAAGTCTCAGGT                 ATAGGTCCTGGCTGAGATTCTATTAATAAAACAAGAGAGAAAGAAGGCGGGCGCCCATTAGA                 ATCTGCTCGGCTGCCAGTAAGTTGCCAGCAAACAGGAACACAAACAAACCAAGGGTGTTGGC                 CCCTACAGGCTCCCAAGGCGGGGGTTGGGCACAGGGCCAAGCTCTGCCACCACAGGAGGCAA                 GTAGACATGCAGGAACACATGGCCCTGGCTAGGAAAGGGAGGAAATAGAGGCCACAGCCAAA                 GTTAGGCTGCCGCCCCACCTGTGTACCCAGGGTGAGAGACCTTGGGAGGTCGTCAGCTTAAC                 CAGCGCCGCCCTCACCCCATTGCCAACTTCCGGCTCACACAAAACCACTCCCAAAAATTGAA                 GACTGGCCAAAATCCAGCTTCCGTCCCCGGGACGTGGTGCTTCCTAAAGGCGGGGCTCATGG                 ATTAGCAGGGGCTTAGTGTGTCATGGTGAGGCAGGCAAGGCGAGCAACGGGGGCTTAGTGGC                 TCAAAGTGATGCATCCCAAAGGCAGCCACCACGCTGGAGGGACATTGTCCACGGGACAAGGC                 ACAGGCCAGGTCATGACCCAGGAAGTGGCGAGCATCGGTCAGCTGACCAAATGTGCAAAGGT                 GACAAGTCAGTAAGGCACGCGGGGGGCCACGTCACCCCGGGGTGCTGGGGTGGGGGATGGGC                 TCAGGCAACCGTAAGGGAGGGGGGGGTAGGGGGGGGAGGGATTACACTATAGGGTTCCCTTC                 CTCTAGGTTCTATATACCTATAGGTATATACCCAGCTGCAATACCCTATTCCACCACTAGGT                 TAATAACCTATAGGTTATTCTACCATTAAAACGGAAGGAGGAAGGGTGGCGCACCTTAAGGT                 AGGGTAGGGGGGTACCCCAGTAGGAACCTAGCTGAATCCTACCTAGCTCCACCCACCTGGTA                 TATAGGGGCGGAGCTTAGGATACCTCCAGGATAATGGAACCCTATGGAGACCTACCTCTAGG                 CTCCACCCACTAGGTATATCGGGGCGGAGCCCACTCCTCCCCCTCCTGGTTCAACCCTATGG                 AGGGGACCCTCCTGAGGCTCCGCCTACCCCAAATCTCGCGGGCCTCTAGCCCCTCCTCCTCT                 CGTTATCCCAATAGAATGACCTCCAGGTACCACCCACCTGGTTACACACCTTAATGTAACCC                 AACGGGCTAAAATCACACACCTGAATTAACCAATGAGAAGCCCCCCACACCTGAGCAAACCT                 TAAGGTATTGCACAGAAACCCCAAAAAGAGGATAAAAGAAGGCGAGCTGGCCCGGCTCGCCA                 GCGTCGTCCAGACGCTCGGGGGGTGCACACCTCCCAGCCGCAAGCTTGGCATTCCGGTACTG                 TTGGTAAAATGGAAGACGCCAAAAACATAAAGAAAGGCCCGGCGCCATTCTATCCTCTAGAG                 GATGGAACCGCTGGAGAGCAACTGCATAAGGCTATGAAGAGATACGCCCTGGTTCCTGGAAC                 AATTGCTTTTACAGATGCACATATCGAGGTGAACATCACGTACGCGGAATACTTCGAAATGT                 CCGTTCGGTTGGCAGAAGCTATGAAACGATATGGGCTGAATACAAATCACAGAATCGTCGTA                 TGCAGTGAAAACTCTCTTCAATTCTTTATGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGC                 AGTTGCGCCCGCGAACGACATTTATAATGAACGTGAATTGCTCAACAGTATGAACATTTCGC                 AGCCTACCGTAGTGTTTGTTTCCAAAAAGGGGTTGCAAAAAATTTTGAACGTGCAAAAAAAA                 TTACCAATAATCCAGAAAATTATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTC                 GATGTACACGTTCGTCACATCTCATCTACCTCCCGGTTTTAATGAATACGATTTTGTACCAG                 AGTCCTTTGATCGTGACAAAACAATTGCACTGATAATGAATTCCTCTGGATCTACTGGGTTA                 CCTAAGGGTGTGGCCCTTCCGCATAGAACTGCCTGCGTCAGATTCTCGCATGCCAGAGATCC                 TATTTTTGGCAATCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACG                 GTTTTGGAATGTTTACTACACTCGGATATTTGATATGTGGATTTCGAGTCGTCTTAATGTAT                 AGATTTGAAGAAGAGCTGTTTTTACGATCCCTTCAGGATTACAAAATTCAAAGTGCGTTGCT                 AGTACCAACCCTATTTTCATTCTTCGCCAAAAGCACTCTGATTGACAAATACGATTTATCTA                 ATTTACACGAAATTGCTTCTGGGGGCGCACCTCTTTCGAAAGAAGTCGGGGAAGCGGTTGCA                 AAACGCTTCCATCTTCCAGGGATACGACAAGGATATGGGCTCACTGAGACTACATCAGCTAT                 TCTGATTACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTTTG                 AAGCGAAGGTTGTGGATCTGGATACCGGGAAAACGCTGGGCGTTAATCAGAGAGGCGAATTA                 TGTGTCAGAGGACCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAACGCCTT                 GATTGACAAGGATGGATGGCTACATTCTGGAGACATAGCTTACTGGGACGAAGACGAACACT                 TCTTCATAGTTGACCGCTTGAAGTCTTTAATTAAATACAAAGGATATCAGGTGGCCCCCGCT                 GAATTGGAATCGATATTGTTACAACACCCCAACATCTTCGACGCGGGCGTGGCAGGTCTTCC                 CGACGATGACGCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAGCACGGAAAGACGATGA                 CGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGCGAAAAAGTTGCGCGGA                 GGAGTTGTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGAAAACTCGACGCAAGAAAAAT                 CAGAGAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGTCCAAATTGTAAAATGTAACTGTAT                 TCAGCGATGACGAAATTCTTAGCTATTGTAATGACTCTAGAGGATCTTTGTGAAGGAACCTT                 ACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAAT                 ATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGAT                 TCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGT                 TTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACT                 CCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTT                 TTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGG                 AAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGG                 CATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGC                 TATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATA                 AGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGA                 GGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATG                 CAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATC                 ACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCAT                 CAATGTATCTTATCATGTCTGGATCCTCTACGCCGGACGCATCGTGGCCGGCATCACCGGCG                 CCACAGGTGCGGTTGCTGGCGCCTATATCGCCGACATCACCGATGGGGAAGATCGGGCTCGC                 CACTTCGGGCTCATGAGCGCTTGTTTCGGCGTGGGTATGGTGGCAGGCCCCGTGGCCGGGGG                 ACTGTTGGGCGCCATCTCCTTGCATGCACCATTCCTTGCGGCGGCGGTGCTCAACGGCCTCA                 ACCTACTACTGGGCTGCTTCCTAATGCAGGAGTCGCATAAGGGAGAGCGTCGACTACGCGAT                 CATGGCGACCACACCCGTCTCGACCGATGCCCTTGAGAGCCTTCAACCCAGTCAGCTCCTTC                 CGGTGGGCGCGGGGCATGACTATCGTCGCCGCACTTATGACTGTCTTCTTTATCATGCAACT                 CGTAGGACAGGTGCCCTGGCCGGGGTCCCGCGGAAACTCGGCCGTGGTGACAGGAAAAGGAC                 AAGCAGCGAAAATTCACGCCCCCTTGGGAGGTGGCGGCATATGCAAAGGATAGCACTCCCAC                 TCTACTACTGGGTATCATATGCTGACTGTATATGCATGAGGATAGCATATGCTACCCGGATA                 CAGATTAGGATAGCATATACTACCCAGATATAGATTAGGATAGCATATGCTACCCAGATATA                 GATTAGGATAGCCTATGCTACCCAGATATAAATTAGGATAGCATATACTACCCAGATATAGA                 TTAGGATAGCATATGCTACCCAGATATAGATTAGGATAGCCTATGCTACCCAGATATAGATT                 AGGATAGCATATGCTACCCAGATATAGATTAGGATAGCATATGCTATCCAGATATTTGGGTA                 GTATATGCTACCCAGATATAAATTAGGATAGCATATACTACCCTAATCTCTATTAGGATAGC                 ATATGCTACCCGGATACAGATTAGGATAGCATATACTACCCAGATATAGATTAGGATAGCAT                 ATGCTACCCAGATATAGATTAGGATAGCCTATGCTACCCAGATATAAATTAGGATAGCATAT                 ACTACCCAGATATAGATTAGGATAGCATATGCTACCCAGATATAGATTAGGATAGCCTATGC                 TACCCAGATATAGATTAGGATAGCATATGCTATCCAGATATTTGGGTAGTATATGCTACCCA                 TGGCAACATTAGCCCACCGTGCTCTCAGCGACCTCGTGAATATGAGGACCAACAACCCTGTG                 CTTGGCGCTCAGGCGCAAGTGTGTGTAATTTGTCCTCCAGATCGCAGCAATCGCGCCCCTAT                 CTTGGCCCGCCCACCTACTTATGCAGGTATTCCCCGGGGTGCCATTAGTGGTTTTGTGGGCA                 AGTGGTTTGACCGCAGTGGTTAGCGGGGTTACAATCAGCCAAGTTATTACACCCTTATTTTA                 CAGTCCAAAACCGCAGGGCGGCGTGTGGGGGCTGACGCGTGCCCCCACTCCACAATTTCAAA                 AAAAAGAGTGGCCACTTGTCTTTGTTTATGGGCCCCATTGGCGTGGAGCCCCGTTTAATTTT                 CGGGGGTGTTAGAGACAACCAGTGGAGTCCGCTGCTGTCGGCGTCCACTCTCTTTCCCCTTG                 TTACAAATAGAGTGTAACAACATGGTTCACCTGTCTTGGTCCCTGCCTGGGACACATCTTAA                 TAACCCCAGTATCATATTGCACTAGGATTATGTGTTGCCCATAGCCATAAATTCGTGTGAGA                 TGGACATCCAGTCTTTACGGCTTGTCCCCACCCCATGGATTTCTATTGTTAAAGATATTCAG                 AATGTTTCATTCCTACACTAGTATTTATTGCCCAAGGGGTTTGTGAGGGTTATATTGGTGTC                 ATAGCACAATGCCACCACTGAACCCCCCGTCCAAATTTTATTCTGGGGGCGTCACCTGAAAC                 CTTGTTTTCGAGCACCTCACATACACCTTACTGTTCACAACTCAGCAGTTATTCTATTAGCT                 AAACGAAGGAGAATGAAGAAGCAGGCGAAGATTCAGGAGAGTTCACTGCCCGCTCCTTGATC                 TTCAGCCACTGCCCTTGTGACTAAAATGGTTCACTACCCTCGTGGAATCCTGACCCCATGTA                 AATAAAACCGTGACAGCTCATGGGGTGGGAGATATCGCTGTTCCTTAGGACCCTTTTACTAA                 CCCTAATTCGATAGCATATGCTTCCCGTTGGGTAACATATGCTATTGAATTAGGGTTAGTCT                 GGATAGTATATACTACTACCCGGGAAGCATATGCTACCCGTTTAGGGTTAACAAGGGGGCCT                 TATAAACACTATTGCTAATGCCCTCTTGAGGGTCCGCTTATCGGTAGCTACACAGGCCCCTC                 TGATTGACGTTGGTGTAGCCTCCCGTAGTCTTCCTGGGCCCCTGGGAGGTACATGTCCCCCA                 GCATTGGTGTAAGAGCTTCAGCCAAGAGTTACACATAAAGGCAATGTTGTGTTGCAGTCCAC                 AGACTGCAAAGTCTGCTCCAGGATGAAAGCCACTCAGTGTTGGCAAATGTGCACATCCATTT                 ATAAGGATGTCAACTACAGTCAGAGAACCCCTTTGTGTTTGGTCCCCCCCCGTGTCACATGT                 GGAACAGGGCCCAGTTGGCAAGTTGTACCAACCAACTGAAGGGATTACATGCACTGCCCCGT                 GACCAATACAAAACAAAAGCGCTCCTCGTACCAGCGAAGAAGGGGCAGAGATGCCGTAGTCA                 GGTTTAGTTCGTCCGGCGGCGCCAGAAATCCGCGCGGTGGTTTTTGGGGGTCGGGGGTGTTT                 GGCAGCCACAGACGCCCGGTGTTCGTGTCGCGCCAGTACATGCGGTCCATGCCCAGGCCATC                 CAAAAACCATGGGTCTGTCTGCTCAGTCCAGTCGTGGACCTGACCCCACGCAACGCCCAAAA                 GAATAACCCCCACGAACCATAAACCATTCCCCATGGGGGACCCCGTCCCTAACCCACGGGGC                 CCGTGGCTATGGCGGGCTTGCCGCCCCGACGTTGGCTGCGAGCCCTGGGCCTTCACCCGAAC                 TTGGGGGTTGGGGTGGGGAAAAGGAAGAAACGCGGGCGTATTGGCCCCAATGGGGTCTCGGT                 GGGGTATCGACAGAGTGCCAGCCCTGGGACCGAACCCCGCGTTTATGAACAAACGACCCAAC                 ACCCGTGCGTTTTATTCTGTCTTTTTATTGCCGTCATAGCGCGGGTTCCTTCCGGTATTGTC                 TCCTTCCGTGTTTCAGTTAGCCTCCCCCATCTCCCGATCCCCTCGGACGAGTGCTGGGGCGT                 CGGTTTCCACTATCGGCGAGTACTTCTACACAGCCATCGGTCCAGACGGCCGCGCTTCTGCG                 GGCGATTTGTGTACGCCCGACAGTCCCGGCTCCGGATCGGACGATTGCGTCGCATCGACCCT                 GCGCCCAAGCTGCATCATCGAAATTGCCGTCAACCAAGCTCTGATAGAGTTGGTCAAGACCA                 ATGCGGAGCATATACGCCCGGAGCCGCGGCGATCCTGCAAGCTCCGGATGCCTCCGCTCGAA                 GTAGCGCGTCTGCTGCTCCATACAAGCCAACCACGGCCTCCAGAAGAAGATGTTGGCGACCT                 CGTATTGGGAATCCCCGAACATCGCCTCGCTCCAGTCAATGACCGCTGTTATGCGGCCATTG                 TCCGTCAGGACATTGTTGGAGCCGAAATCCGCGTGCACGAGGTGCCGGACTTCGGGGCAGTC                 CTCGGCCCAAAGCATCAGCTCATCGAGAGCCTGCGCGACGGACGCACTGACGGTGTCGTCCA                 TCACAGTTTGCCAGTGATACACATGGGGATCAGCAATCGCGCATATGAAATCACGCCATGTA                 GTGTATTGACCGATTCCTTGCGGTCCGAATGGGCCGAACCCGCTCGTCTGGCTAAGATCGGC                 CGCAGCGATCGCATCCATGGCCTCCGCGACCGGCTGCAGAACAGCGGGCAGTTCGGTTTCAG                 GCAGGTCTTGCAACGTGACACCCTGTGCACGGCGGGAGATGCAATAGGTCAGGCTCTCGCTG                 AATTCCCCAATGTCAAGCACTTCCGGAATCGGGAGCGCGGCCGATGCAAAGTGCCGATAAAC                 ATAACGATCTTTGTAGAAACCATCGGCGCAGCTATTTACCCGCAGGACATATCCACGCCCTC                 CTACATCGAAGCTGAAAGCACGAGATTCTTCGCCCTCCGAGAGCTGCATCAGGTCGGAGACG                 CTGTCGAACTTTTCGATCAGAAACTTCTCGACAGACGTCGCGGTGAGTTCAGGCTTTTTCAT                 ATCTCATTGCCCCCGGGGGATCTGCGGCACGCTGTTGACGCTGTTAAGCGGGTCGCTGCAGG                 GTCGCTCGGTGTTCGAGGCCACACGCGTCACCTTAATATGCGAAGTGGACCTGGGACCGCGC                 CGCCCCGACTGCATCTGCGTGTTCGAATTCGCCAATGACAAGACGCTGGGCGGGGTTTGTGT                 CATCATAGAACTAAAGACATGCAAATATATTTCTTCCGGGGACACCGCCAGCAAACGCGAGC                 AACGGGCCACGGGGATGAAGCAGGGCGGCACCTCGCTAACGGATTCACCACTCCAAGAATTG                 GAGCCAATCAATTCTTGCGGAGAACTGTGAATGCGCAAACCAACCCTTGGCAGAACATATCC                 ATCGCGTCCGCCATCTCCAGCAGCCGCACGCGGCGCATCTCGGGCAGGCCGACGCGCTGGGC                 TACGTCTTGCTGGCGTTCGCGACGCGAGGCTGGATGGCCTTCCCCATTATGATTCTTCTCGC                 TTCCGGCGGCATCGGGATGCCCGCGTTGCAGGCCATGCTGTCCAGGCAGGTAGATGACGACC                 ATCAGGGCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGC                 TCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACA                 GGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGAC                 CCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAAT                 GCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCAC                 GAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCC                 GGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGT                 ATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACA                 GTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTG                 ATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGC                 GCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGG                 AACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGAT                 CCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTG                 ACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCC                 ATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCC                 CAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACC                 AGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCT                 ATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGT                 TGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCG                 GTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCC                 TTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGC                 AGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGT                 ACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCA                 ACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTC                 TTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTC                 GTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACA                 GGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACT                 CTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATAT                 TTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCA                 CCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAG                 GCCCTTTCGTCTTCAAGAATTCTCATGTTTGACAGCTTATCATCGATAAGCTTTAATGCGGT                 AGTTTATCACAGTTAAATTGCTAACGCAGTCAGGCACCGTGTATGAAATCTAACAATGCGCT                 CATCGTCATCCTCGGCACCGTCACCCTGGATGCTGTAGGCATAGGCTTGGTTATGCCGGTAC                 TGCCGGGCCTCTTGCGGGATATCGTCCATTCCGACAGCATCGCCAGTCACTATGGCGTGCTG                 CTAGCGCTATATGCGTTGATGCAATTTCTATGCGCACCCGTTCTCGGAGCACTGTCCGACCG                 CTTTGGCCGCCGCCCAGTCCTGCTCGCTTCGCTACTTGGAGCCACTATCGACTACGCGATCA                 TGGCGACCACACCCGTCCTGTG                                                         __________________________________________________________________________

    __________________________________________________________________________     SEQUENCE LISTING                                                               (1) GENERAL INFORMATION:                                                       (iii) NUMBER OF SEQUENCES: 2                                                   (2) INFORMATION FOR SEQ ID NO:1:                                               (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 10580 base pairs                                                   (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: circular                                                         (ii) MOLECULE TYPE: DNA (genomic)                                              (iii) HYPOTHETICAL: NO                                                         (iv) ANTI-SENSE: NO                                                            (x) PUBLICATION INFORMATION:                                                   (H) DOCUMENT NUMBER: WO PCT/DE91/00652                                         (I) FILING DATE: 16-AUG-1991                                                   (J) PUBLICATION DATE: 04-MAR- 1993                                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                        TTCTCATGTTTGACAGCTTATCATCGATAAGCTTTAATGCGGTAGTTTATCACAGTTAAA60                 TTGCTAACGCAGTCAGGCACCGTGTATGAAATCTAACAATGCGCTCATCGTCATCCTCGG120                CACCGTCACCCTGGATGCTGTAGGCATAGGCTTGGTTATGCCGGTACTGCCGGGCCTCTT180                GCGGGATATCGTCCATTCCGACAGCATCGCCAGTCACTATGGCGTGCTGCTAGCGCTATA240                TGCGTTGATGCAATTTCTATGCGCACCCGTTCTCGGAGCACTGTCCGACCGCTTTGGCCG300                CCGCCCAGTCCTGCTCGCTTCGCTACTTGGAGCCACTATCGACTACGCGATCATGGCGAC360                CACACCCGTCCTGTGGATCCTCTACGCCGGACGCATCGTGGCCGGCATCACCGGCGCCAC420                AGGTGCGGTTGCTGGCGCCTATATCGCCGACATCACCGATGGGGAAGATCGGGCTCGCCA480                CTTCGGGCTCATGAGCGCTTGTTTCGGCGTGGGTATGGTGGCAGGCCCCGTGGCCGGGGG540                ACTGTTGGGCGCCATCTCCTTGCATGCCTGCAGGTCGACTCTAGAACTGGATACTCGTGG600                GGGGTAAAGAAGGTGAATAAAAATTACAAACATTCTCTGCCCAGCCTTCAGACTATCAAA660                CCTAGATAATCATTCTATAAATAACCCAGGGCTTGCATCACAATTTAAAGGGAAGTTAAG720                ATGGGGGATTACCTTCTAAGAGGAAATGACAAGCCACAGACATCCCCACATATAGCCGGG780                TGTCCACTGTCTAATGTCAGTAATTTAATTCCATAGTGAAAATAGCACCCCCAACTCAAT840                TTGGAATCCAGAAACTATATTGCACACCAACACCCCCTCCTCTGCACATGAGCAAGACAA900                GACATCTATGTTTATCTCTAAATGTGCCATGGAACCCGGTTGCCCATGCAGTGGTGTCAG960                ACAGGAAAATGGTTAATTAACCACATCTTAAATTGCCACCTTAGGCAAATTGAAAATAGG1020               CAGGCTGAAGAATTGCACAATACCCTAAACAGGTAAGAGGAATTAGTGACATTTTATGAA1080               TTTTTTTACAAACTTTCACACTCAAGAATAGAAACCCAATACCAACAGGTGTGCAGGTGT1140               GCATGACAAATCTTGGGGGTCTCAGAACCCAGGACCAGACTTTGAAGTCTCAGGTATAGG1200               TCCTGGCTGAGATTCTATTAATAAAACAAGAGAGAAAGAAGGCGGGCGCCCATTAGAATC1260               TGCTCGGCTGCCAGTAAGTTGCCAGCAAACAGGAACACAAACAAACCAAGGGTGTTGGCC1320               CCTACAGGCTCCCAAGGCGGGGGTTGGGCACAGGGCCAAGCTCTGCCACCACAGGAGGCA1380               AGTAGACATGCAGGAACACATGGCCCTGGCTAGGAAAGGGAGGAAATAGAGGCCACAGCC1440               AAAGTTAGGCTGCCGCCCCACCTGTGTACCCAGGGTGAGAGACCTTGGGAGGTCGTCAGC1500               TTAACCAGCGCCGCCCTCACCCCATTGCCAACTTCCGGCTCACACAAAACCACTCCCAAA1560               AATTGAAGACTGGCCAAAATCCAGCTTCCGTCCCCGGGACGTGGTGCTTCCTAAAGGCGG1620               GGCTCATGGATTAGCAGGGGCTTAGTGTGTCATGGTGAGGCAGGCAAGGCGAGCAACGGG1680               GGCTTAGTGGCTCAAAGTGATGCATCCCAAAGGCAGCCACCACGCTGGAGGGACATTGTC1740               CACGGGACAAGGCACAGGCCAGGTCATGACCCAGGAAGTGGCGAGCATCGGTCAGCTGAC1800               CAAATGTGCAAAGGTGACAAGTCAGTAAGGCACGCGGGGGGCCACGTCACCCCGGGGTGC1860               TGGGGTGGGGGATGGGCTCAGGCAACCGTAAGGGAGGGGGGGGTAGGGGGGGGAGGGATT1920               ACACTATAGGGTTCCCTTCCTCTAGGTTCTATATACCTATAGGTATATACCCAGCTGCAA1980               TACCCTATTCCACCACTAGGTTAATAACCTATAGGTTATTCTACCATTAAAACGGAAGGA2040               GGAAGGGTGGCGCACCTTAAGGTAGGGTAGGGGGGTACCCCAGTAGGAACCTAGCTGAAT2100               CCTACCTAGCTCCACCCACCTGGTATATAGGGGCGGAGCTTAGGATACCTCCAGGATAAT2160               GGAACCCTATGGAGACCTACCTCTAGGCTCCACCCACTAGGTATATCGGGGCGGAGCCCA2220               CTCCTCCCCCTCCTGGTTCAACCCTATGGAGGGGACCCTCCTGAGGCTCCGCCTACCCCA2280               AATCTCGCGGGCCTCTAGCCCCTCCTCCTCTCGTTATCCCAATAGAATGACCTCCAGGTA2340               CCACCCACCTGGTTACACACCTTAATGTAACCCAACGGGCTAAAATCACACACCTGAATT2400               AACCAATGAGAAGCCCCCCACACCTGAGCAAACCTTAAGGTATTGCACAGAAACCCCAAA2460               AAGAGGATAAAAGAAGGCGAGCTGGCCCGGCTCGCCAGCGTCGTCCAGACGCTCGGGGGG2520               TGCACACCTCCCAGCCGCAAGCTTGGCGAGATTTTCAGGAGCTAAGGAAGCTAAAATGGA2580               GAAAAAAATCACTGGATATACCACCGTTGATATATCCCAATGGCATCGTAAAGAACATTT2640               TGAGGCATTTCAGTCAGTTGCTCAATGTACCTATAACCAGACCGTTCAGCTGGATATTAC2700               GGCCTTTTTAAAGACCGTAAAGAAAAATAAGCACAAGTTTTATCCGGCCTTTATTCACAT2760               TCTTGCCCGCCTGATGAATGCTCATCCGGAATTCCGTATGGCAATGAAAGACGGTGAGCT2820               GGTGATATGGGATAGTGTTCACCCTTGTTACACCGTTTTCCATGAGCAAACTGAAACGTT2880               TTCATCGCTCTGGAGTGAATACCACGACGATTTCCGGCAGTTTCTACACATATATTCGCA2940               AGATGTGGCGTGTTACGGTGAAAACCTGGCCTATTTCCCTAAAGGGTTTATTGAGAATAT3000               GTTTTTCGTCTCAGCCAATCCCTGGGTGAGTTTCACCAGTTTTGATTTAAACGTGGCCAA3060               TATGGACAACTTCTTCGCCCCCGTTTTCACCATGGGCAAATATTATACGCAAGGCGACAA3120               GGTGCTGATGCCGCTGGCGATTCAGGTTCATCATGCCGTCTGTGATGGCTTCCATGTCGG3180               CAGAATGCTTAATGAATTACAACAGTACTGCGATGAGTGGCAGGGCGGGGCGTAATTTTT3240               TTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAG3300               CGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGAC3360               ATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAG3420               TGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGG3480               AACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGA3540               AGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAA3600               AAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAG3660               TCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAA3720               AGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCA3780               TAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGC3840               TATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAA3900               TAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTG3960               TAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAA4020               TGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCA4080               ATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGT4140               CCAAACTCATCAATGTATCTTATCATGTCTGGATCCCCGGGTACCGAGCTCGCCTCGACC4200               GATGCCCTTGAGAGCCTTCAACCCAGTCAGCTCCTTCCGGTGGGCGCGGGGCATGACTAT4260               CGTCGCCGCACTTATGACTGTCTTCTTTATCATGCAACTCGTAGGACAGGTGCCCTGGCC4320               GGGGTCCCGCGGAAACTCGGCCGTGGTGACAGGAAAAGGACAAGCAGCGAAAATTCACGC4380               CCCCTTGGGAGGTGGCGGCATATGCAAAGGATAGCACTCCCACTCTACTACTGGGTATCA4440               TATGCTGACTGTATATGCATGAGGATAGCATATGCTACCCGGATACAGATTAGGATAGCA4500               TATACTACCCAGATATAGATTAGGATAGCATATGCTACCCAGATATAGATTAGGATAGCC4560               TATGCTACCCAGATATAAATTAGGATAGCATATACTACCCAGATATAGATTAGGATAGCA4620               TATGCTACCCAGATATAGATTAGGATAGCCTATGCTACCCAGATATAGATTAGGATAGCA4680               TATGCTACCCAGATATAGATTAGGATAGCATATGCTATCCAGATATTTGGGTAGTATATG4740               CTACCCAGATATAAATTAGGATAGCATATACTACCCTAATCTCTATTAGGATAGCATATG4800               CTACCCGGATACAGATTAGGATAGCATATACTACCCAGATATAGATTAGGATAGCATATG4860               CTACCCAGATATAGATTAGGATAGCCTATGCTACCCAGATATAAATTAGGATAGCATATA4920               CTACCCAGATATAGATTAGGATAGCATATGCTACCCAGATATAGATTAGGATAGCCTATG4980               CTACCCAGATATAGATTAGGATAGCATATGCTATCCAGATATTTGGGTAGTATATGCTAC5040               CCATGGCAACATTAGCCCACCGTGCTCTCAGCGACCTCGTGAATATGAGGACCAACAACC5100               CTGTGCTTGGCGCTCAGGCGCAAGTGTGTGTAATTTGTCCTCCAGATCGCAGCAATCGCG5160               CCCCTATCTTGGCCCGCCCACCTACTTATGCAGGTATTCCCCGGGGTGCCATTAGTGGTT5220               TTGTGGGCAAGTGGTTTGACCGCAGTGGTTAGCGGGGTTACAATCAGCCAAGTTATTACA5280               CCCTTATTTTACAGTCCAAAACCGCAGGGCGGCGTGTGGGGGCTGACGCGTGCCCCCACT5340               CCACAATTTCAAAAAAAAGAGTGGCCACTTGTCTTTGTTTATGGGCCCCATTGGCGTGGA5400               GCCCCGTTTAATTTTCGGGGGTGTTAGAGACAACCAGTGGAGTCCGCTGCTGTCGGCGTC5460               CACTCTCTTTCCCCTTGTTACAAATAGAGTGTAACAACATGGTTCACCTGTCTTGGTCCC5520               TGCCTGGGACACATCTTAATAACCCCAGTATCATATTGCACTAGGATTATGTGTTGCCCA5580               TAGCCATAAATTCGTGTGAGATGGACATCCAGTCTTTACGGCTTGTCCCCACCCCATGGA5640               TTTCTATTGTTAAAGATATTCAGAATGTTTCATTCCTACACTAGTATTTATTGCCCAAGG5700               GGTTTGTGAGGGTTATATTGGTGTCATAGCACAATGCCACCACTGAACCCCCCGTCCAAA5760               TTTTATTCTGGGGGCGTCACCTGAAACCTTGTTTTCGAGCACCTCACATACACCTTACTG5820               TTCACAACTCAGCAGTTATTCTATTAGCTAAACGAAGGAGAATGAAGAAGCAGGCGAAGA5880               TTCAGGAGAGTTCACTGCCCGCTCCTTGATCTTCAGCCACTGCCCTTGTGACTAAAATGG5940               TTCACTACCCTCGTGGAATCCTGACCCCATGTAAATAAAACCGTGACAGCTCATGGGGTG6000               GGAGATATCGCTGTTCCTTAGGACCCTTTTACTAACCCTAATTCGATAGCATATGCTTCC6060               CGTTGGGTAACATATGCTATTGAATTAGGGTTAGTCTGGATAGTATATACTACTACCCGG6120               GAAGCATATGCTACCCGTTTAGGGTTAACAAGGGGGCCTTATAAACACTATTGCTAATGC6180               CCTCTTGAGGGTCCGCTTATCGGTAGCTACACAGGCCCCTCTGATTGACGTTGGTGTAGC6240               CTCCCGTAGTCTTCCTGGGCCCCTGGGAGGTACATGTCCCCCAGCATTGGTGTAAGAGCT6300               TCAGCCAAGAGTTACACATAAAGGCAATGTTGTGTTGCAGTCCACAGACTGCAAAGTCTG6360               CTCCAGGATGAAAGCCACTCAGTGTTGGCAAATGTGCACATCCATTTATAAGGATGTCAA6420               CTACAGTCAGAGAACCCCTTTGTGTTTGGTCCCCCCCCGTGTCACATGTGGAACAGGGCC6480               CAGTTGGCAAGTTGTACCAACCAACTGAAGGGATTACATGCACTGCCCCGTGACCAATAC6540               AAAACAAAAGCGCTCCTCGTACCAGCGAAGAAGGGGCAGAGATGCCGTAGTCAGGTTTAG6600               TTCGTCCGGCGGCGCCAGAAATCCGCGCGGTGGTTTTTGGGGGTCGGGGGTGTTTGGCAG6660               CCACAGACGCCCGGTGTTCGTGTCGCGCCAGTACATGCGGTCCATGCCCAGGCCATCCAA6720               AAACCATGGGTCTGTCTGCTCAGTCCAGTCGTGGACCTGACCCCACGCAACGCCCAAAAG6780               AATAACCCCCACGAACCATAAACCATTCCCCATGGGGGACCCCGTCCCTAACCCACGGGG6840               CCCGTGGCTATGGCGGGCTTGCCGCCCCGACGTTGGCTGCGAGCCCTGGGCCTTCACCCG6900               AACTTGGGGGTTGGGGTGGGGAAAAGGAAGAAACGCGGGCGTATTGGCCCCAATGGGGTC6960               TCGGTGGGGTATCGACAGAGTGCCAGCCCTGGGACCGAACCCCGCGTTTATGAACAAACG7020               ACCCAACACCCGTGCGTTTTATTCTGTCTTTTTATTGCCGTCATAGCGCGGGTTCCTTCC7080               GGTATTGTCTCCTTCCGTGTTTCAGTTAGCCTCCCCCATCTCCCGATCCCCTCGGACGAG7140               TGCTGGGGCGTCGGTTTCCACTATCGGCGAGTACTTCTACACAGCCATCGGTCCAGACGG7200               CCGCGCTTCTGCGGGCGATTTGTGTACGCCCGACAGTCCCGGCTCCGGATCGGACGATTG7260               CGTCGCATCGACCCTGCGCCCAAGCTGCATCATCGAAATTGCCGTCAACCAAGCTCTGAT7320               AGAGTTGGTCAAGACCAATGCGGAGCATATACGCCCGGAGCCGCGGCGATCCTGCAAGCT7380               CCGGATGCCTCCGCTCGAAGTAGCGCGTCTGCTGCTCCATACAAGCCAACCACGGCCTCC7440               AGAAGAAGATGTTGGCGACCTCGTATTGGGAATCCCCGAACATCGCCTCGCTCCAGTCAA7500               TGACCGCTGTTATGCGGCCATTGTCCGTCAGGACATTGTTGGAGCCGAAATCCGCGTGCA7560               CGAGGTGCCGGACTTCGGGGCAGTCCTCGGCCCAAAGCATCAGCTCATCGAGAGCCTGCG7620               CGACGGACGCACTGACGGTGTCGTCCATCACAGTTTGCCAGTGATACACATGGGGATCAG7680               CAATCGCGCATATGAAATCACGCCATGTAGTGTATTGACCGATTCCTTGCGGTCCGAATG7740               GGCCGAACCCGCTCGTCTGGCTAAGATCGGCCGCAGCGATCGCATCCATGGCCTCCGCGA7800               CCGGCTGCAGAACAGCGGGCAGTTCGGTTTCAGGCAGGTCTTGCAACGTGACACCCTGTG7860               CACGGCGGGAGATGCAATAGGTCAGGCTCTCGCTGAATTCCCCAATGTCAAGCACTTCCG7920               GAATCGGGAGCGCGGCCGATGCAAAGTGCCGATAAACATAACGATCTTTGTAGAAACCAT7980               CGGCGCAGCTATTTACCCGCAGGACATATCCACGCCCTCCTACATCGAAGCTGAAAGCAC8040               GAGATTCTTCGCCCTCCGAGAGCTGCATCAGGTCGGAGACGCTGTCGAACTTTTCGATCA8100               GAAACTTCTCGACAGACGTCGCGGTGAGTTCAGGCTTTTTCATATCTCATTGCCCCCGGG8160               GGATCTGCGGCACGCTGTTGACGCTGTTAAGCGGGTCGCTGCAGGGTCGCTCGGTGTTCG8220               AGGCCACACGCGTCACCTTAATATGCGAAGTGGACCTGGGACCGCGCCGCCCCGACTGCA8280               TCTGCGTGTTCGAATTCGCCAATGACAAGACGCTGGGCGGGGTTTGTGTCATCATAGAAC8340               TAAAGACATGCAAATATATTTCTTCCGGGGACACCGCCAGCAAACGCGAGCAACGGGCCA8400               CGGGGATGAAGCAGGGCGGCACCTCGCTAACGGATTCACCACTCCAAGAATTGGAGCCAA8460               TCAATTCTTGCGGAGAACTGTGAATGCGCAAACCAACCCTTGGCAGAACATATCCATCGC8520               GTCCGCCATCTCCAGCAGCCGCACGCGGCGCATCTCGGGCAGGCCGACGCGCTGGGCTAC8580               GTCTTGCTGGCGTTCGCGACGCGAGGCTGGATGGCCTTCCCCATTATGATTCTTCTCGCT8640               TCCGGCGGCATCGGGATGCCCGCGTTGCAGGCCATGCTGTCCAGGCAGGTAGATGACGAC8700               CATCAGGGCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATA8760               GGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACC8820               CGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTG8880               TTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGC8940               TTTCTCAATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGG9000               GCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTC9060               TTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGA9120               TTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACG9180               GCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAA9240               AAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTG9300               TTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTT9360               CTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGAT9420               TATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCT9480               AAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTA9540               TCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAA9600               CTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCAC9660               GCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAA9720               GTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAG9780               TAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGG9840               TGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAG9900               TTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTG9960               TCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTC10020              TTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCAT10080              TCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATA10140              CCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAA10200              AACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCA10260              ACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGC10320              AAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCC10380              TTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTG10440              AATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCAC10500              CTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGA10560              GGCCCTTTCGTCTTCAAGAA10580                                                      (2) INFORMATION FOR SEQ ID NO:2:                                               (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 11616 base pairs                                                   (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: circular                                                         (ii) MOLECULE TYPE: DNA (genomic)                                              (iii) HYPOTHETICAL: NO                                                         (iv) ANTI-SENSE: NO                                                            (x) PUBLICATION INFORMATION:                                                   (H) DOCUMENT NUMBER: WO PCT/DE91/00652                                         (I) FILING DATE: 16-AUG-1991                                                   (J) PUBLICATION DATE: 04-MAR- 1993                                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                        GATCTCTAGAACTGGATACTCGTGGGGGGTAAAGAAGGTGAATAAAAATTACAAACATTC60                 TCTGCCCAGCCTTCAGACTATCAAACCTAGATAATCATTCTATAAATAACCCAGGGCTTG120                CATCACAATTTAAAGGGAAGTTAAGATGGGGGATTACCTTCTAAGAGGAAATGACAAGCC180                ACAGACATCCCCACATATAGCCGGGTGTCCACTGTCTAATGTCAGTAATTTAATTCCATA240                GTGAAAATAGCACCCCCAACTCAATTTGGAATCCAGAAACTATATTGCACACCAACACCC300                CCTCCTCTGCACATGAGCAAGACAAGACATCTATGTTTATCTCTAAATGTGCCATGGAAC360                CCGGTTGCCCATGCAGTGGTGTCAGACAGGAAAATGGTTAATTAACCACATCTTAAATTG420                CCACCTTAGGCAAATTGAAAATAGGCAGGCTGAAGAATTGCACAATACCCTAAACAGGTA480                AGAGGAATTAGTGACATTTTATGAATTTTTTTACAAACTTTCACACTCAAGAATAGAAAC540                CCAATACCAACAGGTGTGCAGGTGTGCATGACAAATCTTGGGGGTCTCAGAACCCAGGAC600                CAGACTTTGAAGTCTCAGGTATAGGTCCTGGCTGAGATTCTATTAATAAAACAAGAGAGA660                AAGAAGGCGGGCGCCCATTAGAATCTGCTCGGCTGCCAGTAAGTTGCCAGCAAACAGGAA720                CACAAACAAACCAAGGGTGTTGGCCCCTACAGGCTCCCAAGGCGGGGGTTGGGCACAGGG780                CCAAGCTCTGCCACCACAGGAGGCAAGTAGACATGCAGGAACACATGGCCCTGGCTAGGA840                AAGGGAGGAAATAGAGGCCACAGCCAAAGTTAGGCTGCCGCCCCACCTGTGTACCCAGGG900                TGAGAGACCTTGGGAGGTCGTCAGCTTAACCAGCGCCGCCCTCACCCCATTGCCAACTTC960                CGGCTCACACAAAACCACTCCCAAAAATTGAAGACTGGCCAAAATCCAGCTTCCGTCCCC1020               GGGACGTGGTGCTTCCTAAAGGCGGGGCTCATGGATTAGCAGGGGCTTAGTGTGTCATGG1080               TGAGGCAGGCAAGGCGAGCAACGGGGGCTTAGTGGCTCAAAGTGATGCATCCCAAAGGCA1140               GCCACCACGCTGGAGGGACATTGTCCACGGGACAAGGCACAGGCCAGGTCATGACCCAGG1200               AAGTGGCGAGCATCGGTCAGCTGACCAAATGTGCAAAGGTGACAAGTCAGTAAGGCACGC1260               GGGGGGCCACGTCACCCCGGGGTGCTGGGGTGGGGGATGGGCTCAGGCAACCGTAAGGGA1320               GGGGGGGGTAGGGGGGGGAGGGATTACACTATAGGGTTCCCTTCCTCTAGGTTCTATATA1380               CCTATAGGTATATACCCAGCTGCAATACCCTATTCCACCACTAGGTTAATAACCTATAGG1440               TTATTCTACCATTAAAACGGAAGGAGGAAGGGTGGCGCACCTTAAGGTAGGGTAGGGGGG1500               TACCCCAGTAGGAACCTAGCTGAATCCTACCTAGCTCCACCCACCTGGTATATAGGGGCG1560               GAGCTTAGGATACCTCCAGGATAATGGAACCCTATGGAGACCTACCTCTAGGCTCCACCC1620               ACTAGGTATATCGGGGCGGAGCCCACTCCTCCCCCTCCTGGTTCAACCCTATGGAGGGGA1680               CCCTCCTGAGGCTCCGCCTACCCCAAATCTCGCGGGCCTCTAGCCCCTCCTCCTCTCGTT1740               ATCCCAATAGAATGACCTCCAGGTACCACCCACCTGGTTACACACCTTAATGTAACCCAA1800               CGGGCTAAAATCACACACCTGAATTAACCAATGAGAAGCCCCCCACACCTGAGCAAACCT1860               TAAGGTATTGCACAGAAACCCCAAAAAGAGGATAAAAGAAGGCGAGCTGGCCCGGCTCGC1920               CAGCGTCGTCCAGACGCTCGGGGGGTGCACACCTCCCAGCCGCAAGCTTGGCATTCCGGT1980               ACTGTTGGTAAAATGGAAGACGCCAAAAACATAAAGAAAGGCCCGGCGCCATTCTATCCT2040               CTAGAGGATGGAACCGCTGGAGAGCAACTGCATAAGGCTATGAAGAGATACGCCCTGGTT2100               CCTGGAACAATTGCTTTTACAGATGCACATATCGAGGTGAACATCACGTACGCGGAATAC2160               TTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAACGATATGGGCTGAATACAAATCAC2220               AGAATCGTCGTATGCAGTGAAAACTCTCTTCAATTCTTTATGCCGGTGTTGGGCGCGTTA2280               TTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATTTATAATGAACGTGAATTGCTCAAC2340               AGTATGAACATTTCGCAGCCTACCGTAGTGTTTGTTTCCAAAAAGGGGTTGCAAAAAATT2400               TTGAACGTGCAAAAAAAATTACCAATAATCCAGAAAATTATTATCATGGATTCTAAAACG2460               GATTACCAGGGATTTCAGTCGATGTACACGTTCGTCACATCTCATCTACCTCCCGGTTTT2520               AATGAATACGATTTTGTACCAGAGTCCTTTGATCGTGACAAAACAATTGCACTGATAATG2580               AATTCCTCTGGATCTACTGGGTTACCTAAGGGTGTGGCCCTTCCGCATAGAACTGCCTGC2640               GTCAGATTCTCGCATGCCAGAGATCCTATTTTTGGCAATCAAATCATTCCGGATACTGCG2700               ATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGAATGTTTACTACACTCGGATATTTG2760               ATATGTGGATTTCGAGTCGTCTTAATGTATAGATTTGAAGAAGAGCTGTTTTTACGATCC2820               CTTCAGGATTACAAAATTCAAAGTGCGTTGCTAGTACCAACCCTATTTTCATTCTTCGCC2880               AAAAGCACTCTGATTGACAAATACGATTTATCTAATTTACACGAAATTGCTTCTGGGGGC2940               GCACCTCTTTCGAAAGAAGTCGGGGAAGCGGTTGCAAAACGCTTCCATCTTCCAGGGATA3000               CGACAAGGATATGGGCTCACTGAGACTACATCAGCTATTCTGATTACACCCGAGGGGGAT3060               GATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTTTGAAGCGAAGGTTGTGGATCTG3120               GATACCGGGAAAACGCTGGGCGTTAATCAGAGAGGCGAATTATGTGTCAGAGGACCTATG3180               ATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAACGCCTTGATTGACAAGGATGGA3240               TGGCTACATTCTGGAGACATAGCTTACTGGGACGAAGACGAACACTTCTTCATAGTTGAC3300               CGCTTGAAGTCTTTAATTAAATACAAAGGATATCAGGTGGCCCCCGCTGAATTGGAATCG3360               ATATTGTTACAACACCCCAACATCTTCGACGCGGGCGTGGCAGGTCTTCCCGACGATGAC3420               GCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAGCACGGAAAGACGATGACGGAAAAA3480               GAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGCGAAAAAGTTGCGCGGAGGAGTT3540               GTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGAAAACTCGACGCAAGAAAAATCAGA3600               GAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGTCCAAATTGTAAAATGTAACTGTATTC3660               AGCGATGACGAAATTCTTAGCTATTGTAATGACTCTAGAGGATCTTTGTGAAGGAACCTT3720               ACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAA3780               ATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTT3840               AGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAA3900               ACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAAC3960               ATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAAT4020               TGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTT4080               ACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAA4140               CCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACA4200               GGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAA4260               TTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATC4320               AGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTG4380               AACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAAT4440               GGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAT4500               TCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGATCCTCTACGCC4560               GGACGCATCGTGGCCGGCATCACCGGCGCCACAGGTGCGGTTGCTGGCGCCTATATCGCC4620               GACATCACCGATGGGGAAGATCGGGCTCGCCACTTCGGGCTCATGAGCGCTTGTTTCGGC4680               GTGGGTATGGTGGCAGGCCCCGTGGCCGGGGGACTGTTGGGCGCCATCTCCTTGCATGCA4740               CCATTCCTTGCGGCGGCGGTGCTCAACGGCCTCAACCTACTACTGGGCTGCTTCCTAATG4800               CAGGAGTCGCATAAGGGAGAGCGTCGACTACGCGATCATGGCGACCACACCCGTCTCGAC4860               CGATGCCCTTGAGAGCCTTCAACCCAGTCAGCTCCTTCCGGTGGGCGCGGGGCATGACTA4920               TCGTCGCCGCACTTATGACTGTCTTCTTTATCATGCAACTCGTAGGACAGGTGCCCTGGC4980               CGGGGTCCCGCGGAAACTCGGCCGTGGTGACAGGAAAAGGACAAGCAGCGAAAATTCACG5040               CCCCCTTGGGAGGTGGCGGCATATGCAAAGGATAGCACTCCCACTCTACTACTGGGTATC5100               ATATGCTGACTGTATATGCATGAGGATAGCATATGCTACCCGGATACAGATTAGGATAGC5160               ATATACTACCCAGATATAGATTAGGATAGCATATGCTACCCAGATATAGATTAGGATAGC5220               CTATGCTACCCAGATATAAATTAGGATAGCATATACTACCCAGATATAGATTAGGATAGC5280               ATATGCTACCCAGATATAGATTAGGATAGCCTATGCTACCCAGATATAGATTAGGATAGC5340               ATATGCTACCCAGATATAGATTAGGATAGCATATGCTATCCAGATATTTGGGTAGTATAT5400               GCTACCCAGATATAAATTAGGATAGCATATACTACCCTAATCTCTATTAGGATAGCATAT5460               GCTACCCGGATACAGATTAGGATAGCATATACTACCCAGATATAGATTAGGATAGCATAT5520               GCTACCCAGATATAGATTAGGATAGCCTATGCTACCCAGATATAAATTAGGATAGCATAT5580               ACTACCCAGATATAGATTAGGATAGCATATGCTACCCAGATATAGATTAGGATAGCCTAT5640               GCTACCCAGATATAGATTAGGATAGCATATGCTATCCAGATATTTGGGTAGTATATGCTA5700               CCCATGGCAACATTAGCCCACCGTGCTCTCAGCGACCTCGTGAATATGAGGACCAACAAC5760               CCTGTGCTTGGCGCTCAGGCGCAAGTGTGTGTAATTTGTCCTCCAGATCGCAGCAATCGC5820               GCCCCTATCTTGGCCCGCCCACCTACTTATGCAGGTATTCCCCGGGGTGCCATTAGTGGT5880               TTTGTGGGCAAGTGGTTTGACCGCAGTGGTTAGCGGGGTTACAATCAGCCAAGTTATTAC5940               ACCCTTATTTTACAGTCCAAAACCGCAGGGCGGCGTGTGGGGGCTGACGCGTGCCCCCAC6000               TCCACAATTTCAAAAAAAAGAGTGGCCACTTGTCTTTGTTTATGGGCCCCATTGGCGTGG6060               AGCCCCGTTTAATTTTCGGGGGTGTTAGAGACAACCAGTGGAGTCCGCTGCTGTCGGCGT6120               CCACTCTCTTTCCCCTTGTTACAAATAGAGTGTAACAACATGGTTCACCTGTCTTGGTCC6180               CTGCCTGGGACACATCTTAATAACCCCAGTATCATATTGCACTAGGATTATGTGTTGCCC6240               ATAGCCATAAATTCGTGTGAGATGGACATCCAGTCTTTACGGCTTGTCCCCACCCCATGG6300               ATTTCTATTGTTAAAGATATTCAGAATGTTTCATTCCTACACTAGTATTTATTGCCCAAG6360               GGGTTTGTGAGGGTTATATTGGTGTCATAGCACAATGCCACCACTGAACCCCCCGTCCAA6420               ATTTTATTCTGGGGGCGTCACCTGAAACCTTGTTTTCGAGCACCTCACATACACCTTACT6480               GTTCACAACTCAGCAGTTATTCTATTAGCTAAACGAAGGAGAATGAAGAAGCAGGCGAAG6540               ATTCAGGAGAGTTCACTGCCCGCTCCTTGATCTTCAGCCACTGCCCTTGTGACTAAAATG6600               GTTCACTACCCTCGTGGAATCCTGACCCCATGTAAATAAAACCGTGACAGCTCATGGGGT6660               GGGAGATATCGCTGTTCCTTAGGACCCTTTTACTAACCCTAATTCGATAGCATATGCTTC6720               CCGTTGGGTAACATATGCTATTGAATTAGGGTTAGTCTGGATAGTATATACTACTACCCG6780               GGAAGCATATGCTACCCGTTTAGGGTTAACAAGGGGGCCTTATAAACACTATTGCTAATG6840               CCCTCTTGAGGGTCCGCTTATCGGTAGCTACACAGGCCCCTCTGATTGACGTTGGTGTAG6900               CCTCCCGTAGTCTTCCTGGGCCCCTGGGAGGTACATGTCCCCCAGCATTGGTGTAAGAGC6960               TTCAGCCAAGAGTTACACATAAAGGCAATGTTGTGTTGCAGTCCACAGACTGCAAAGTCT7020               GCTCCAGGATGAAAGCCACTCAGTGTTGGCAAATGTGCACATCCATTTATAAGGATGTCA7080               ACTACAGTCAGAGAACCCCTTTGTGTTTGGTCCCCCCCCGTGTCACATGTGGAACAGGGC7140               CCAGTTGGCAAGTTGTACCAACCAACTGAAGGGATTACATGCACTGCCCCGTGACCAATA7200               CAAAACAAAAGCGCTCCTCGTACCAGCGAAGAAGGGGCAGAGATGCCGTAGTCAGGTTTA7260               GTTCGTCCGGCGGCGCCAGAAATCCGCGCGGTGGTTTTTGGGGGTCGGGGGTGTTTGGCA7320               GCCACAGACGCCCGGTGTTCGTGTCGCGCCAGTACATGCGGTCCATGCCCAGGCCATCCA7380               AAAACCATGGGTCTGTCTGCTCAGTCCAGTCGTGGACCTGACCCCACGCAACGCCCAAAA7440               GAATAACCCCCACGAACCATAAACCATTCCCCATGGGGGACCCCGTCCCTAACCCACGGG7500               GCCCGTGGCTATGGCGGGCTTGCCGCCCCGACGTTGGCTGCGAGCCCTGGGCCTTCACCC7560               GAACTTGGGGGTTGGGGTGGGGAAAAGGAAGAAACGCGGGCGTATTGGCCCCAATGGGGT7620               CTCGGTGGGGTATCGACAGAGTGCCAGCCCTGGGACCGAACCCCGCGTTTATGAACAAAC7680               GACCCAACACCCGTGCGTTTTATTCTGTCTTTTTATTGCCGTCATAGCGCGGGTTCCTTC7740               CGGTATTGTCTCCTTCCGTGTTTCAGTTAGCCTCCCCCATCTCCCGATCCCCTCGGACGA7800               GTGCTGGGGCGTCGGTTTCCACTATCGGCGAGTACTTCTACACAGCCATCGGTCCAGACG7860               GCCGCGCTTCTGCGGGCGATTTGTGTACGCCCGACAGTCCCGGCTCCGGATCGGACGATT7920               GCGTCGCATCGACCCTGCGCCCAAGCTGCATCATCGAAATTGCCGTCAACCAAGCTCTGA7980               TAGAGTTGGTCAAGACCAATGCGGAGCATATACGCCCGGAGCCGCGGCGATCCTGCAAGC8040               TCCGGATGCCTCCGCTCGAAGTAGCGCGTCTGCTGCTCCATACAAGCCAACCACGGCCTC8100               CAGAAGAAGATGTTGGCGACCTCGTATTGGGAATCCCCGAACATCGCCTCGCTCCAGTCA8160               ATGACCGCTGTTATGCGGCCATTGTCCGTCAGGACATTGTTGGAGCCGAAATCCGCGTGC8220               ACGAGGTGCCGGACTTCGGGGCAGTCCTCGGCCCAAAGCATCAGCTCATCGAGAGCCTGC8280               GCGACGGACGCACTGACGGTGTCGTCCATCACAGTTTGCCAGTGATACACATGGGGATCA8340               GCAATCGCGCATATGAAATCACGCCATGTAGTGTATTGACCGATTCCTTGCGGTCCGAAT8400               GGGCCGAACCCGCTCGTCTGGCTAAGATCGGCCGCAGCGATCGCATCCATGGCCTCCGCG8460               ACCGGCTGCAGAACAGCGGGCAGTTCGGTTTCAGGCAGGTCTTGCAACGTGACACCCTGT8520               GCACGGCGGGAGATGCAATAGGTCAGGCTCTCGCTGAATTCCCCAATGTCAAGCACTTCC8580               GGAATCGGGAGCGCGGCCGATGCAAAGTGCCGATAAACATAACGATCTTTGTAGAAACCA8640               TCGGCGCAGCTATTTACCCGCAGGACATATCCACGCCCTCCTACATCGAAGCTGAAAGCA8700               CGAGATTCTTCGCCCTCCGAGAGCTGCATCAGGTCGGAGACGCTGTCGAACTTTTCGATC8760               AGAAACTTCTCGACAGACGTCGCGGTGAGTTCAGGCTTTTTCATATCTCATTGCCCCCGG8820               GGGATCTGCGGCACGCTGTTGACGCTGTTAAGCGGGTCGCTGCAGGGTCGCTCGGTGTTC8880               GAGGCCACACGCGTCACCTTAATATGCGAAGTGGACCTGGGACCGCGCCGCCCCGACTGC8940               ATCTGCGTGTTCGAATTCGCCAATGACAAGACGCTGGGCGGGGTTTGTGTCATCATAGAA9000               CTAAAGACATGCAAATATATTTCTTCCGGGGACACCGCCAGCAAACGCGAGCAACGGGCC9060               ACGGGGATGAAGCAGGGCGGCACCTCGCTAACGGATTCACCACTCCAAGAATTGGAGCCA9120               ATCAATTCTTGCGGAGAACTGTGAATGCGCAAACCAACCCTTGGCAGAACATATCCATCG9180               CGTCCGCCATCTCCAGCAGCCGCACGCGGCGCATCTCGGGCAGGCCGACGCGCTGGGCTA9240               CGTCTTGCTGGCGTTCGCGACGCGAGGCTGGATGGCCTTCCCCATTATGATTCTTCTCGC9300               TTCCGGCGGCATCGGGATGCCCGCGTTGCAGGCCATGCTGTCCAGGCAGGTAGATGACGA9360               CCATCAGGGCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCAT9420               AGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAAC9480               CCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCT9540               GTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCG9600               CTTTCTCAATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTG9660               GGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGT9720               CTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGG9780               ATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTAC9840               GGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGA9900               AAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTT9960               GTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTT10020              TCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGA10080              TTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATC10140              TAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCT10200              ATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATA10260              ACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCA10320              CGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGA10380              AGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGA10440              GTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTG10500              GTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGA10560              GTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTT10620              GTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCT10680              CTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCA10740              TTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAAT10800              ACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGA10860              AAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCC10920              AACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGG10980              CAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTC11040              CTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTT11100              GAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCA11160              CCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACG11220              AGGCCCTTTCGTCTTCAAGAATTCTCATGTTTGACAGCTTATCATCGATAAGCTTTAATG11280              CGGTAGTTTATCACAGTTAAATTGCTAACGCAGTCAGGCACCGTGTATGAAATCTAACAA11340              TGCGCTCATCGTCATCCTCGGCACCGTCACCCTGGATGCTGTAGGCATAGGCTTGGTTAT11400              GCCGGTACTGCCGGGCCTCTTGCGGGATATCGTCCATTCCGACAGCATCGCCAGTCACTA11460              TGGCGTGCTGCTAGCGCTATATGCGTTGATGCAATTTCTATGCGCACCCGTTCTCGGAGC11520              ACTGTCCGACCGCTTTGGCCGCCGCCCAGTCCTGCTCGCTTCGCTACTTGGAGCCACTAT11580              CGACTACGCGATCATGGCGACCACACCCGTCCTGTG11616                                      __________________________________________________________________________ 

We claim:
 1. A method for the in vitro detection of tumor promoters, comprising the following steps:(a) contacting EBV-containing cells which contain pHEBO-DR-CAT or pHEBO-DR-LUC with the substance to be examined; (b) incubating the cells in contact with the substance to be examined in a CO₂ -containing atmosphere at 37° C. for approximately one to three days; (c) determining the quantity of the expressed reporter gene product; and (d) comparing the quantity of the expressed reporter gene to the quantity of the expressed reporter gene from untreated cells.
 2. A DNA construct comprising:(a) the control region for the activation of the Epstein-Barr virus (EBV) DR gene; (b) the episomal origin of replication of EBV (oriP) (c) a reporter gene; (d) a resistance gene R1 for the selection of transfected eukaryotic cells; and (e) a resistance gene R2 for selection in procaryotic cells.
 3. A DNA construct according to claim 2 wherein R2 is an ampicillin resistance gene.
 4. A DNA construct according to claim 2, wherein the reporter gene is chloramphenicol acetyltransferase (CAT).
 5. A DNA construct according to claim 2 wherein the reporter gene is firefly luciferase (LUC).
 6. A DNA construct according to claim 2 wherein R1 is a hygromycin resistance gene.
 7. An in-vitro test for the detection of tumor promoters, comprising the following method steps:(a) EBV-containing cells containing a DNA construct according to claim 2 are brought in contact with the substance to be examined; (b) the cells in contact with the substance to be examined are kept in a CO₂ -containing atmosphere at 37° C. for approximately one to three days; and (c) the quantity of the product expressed from the reporter gene is determined.
 8. The in-vitro test according to claim 7, wherein Raji cells are employed as the EBV-containing cells.
 9. The in-vitro test according to claim 7, wherein the reporter gene is chloramphenicol acetyltransferase (CAT).
 10. The in-vitro test according to claim 7, wherein the reporter gene is firefly luciferase (LUC).
 11. The in-vitro test according to claim 9, wherein the number of EBV-containing cells containing the DNA construct is from 1 to 5×10⁶ per experiment.
 12. The in-vitro test according to claim 10, wherein the number of EBV-containing cells containing the DNA construct is from 0.1 to 1.0×10⁴ cells per experiment.
 13. The in-vitro test according to claim 7, comprising the additional step of:(d) the quantity of the product expressed from the reporter gene is compared to the quantity of the product expressed from the reporter gene from untreated cells. 